Il-17 antibody formulation

ABSTRACT

The present invention provides pharmaceutical formulations for anti-IL-17 antibodies. These anti-IL-17 antibody pharmaceutical formulations can be used to treat rheumatoid arthritis, psoriasis, ankylosing psoriatic arthritis or multiple myeloma.

The present invention relates to the field of medicine. Moreparticularly, the present invention relates to a pharmaceuticalformulation of an anti-IL-17 antibody. This anti-IL-17 antibodypharmaceutical formulation is expected to be useful in treatingRheumatoid Arthritis (RA), Psoriasis (Ps), Ankylosing Spondilitis (AS),Psoriatic Arthritis (PA) or Multiple Myeloma (MM).

Pharmaceutical formulations of anti-IL-17 antibodies are needed for thetreatment of patients with RA, Ps, AS, PA or MM. Certain concentrationsof anti-IL-17 antibodies are needed for pharmaceutical formulations sothat the antibody can be delivered subcutaneously to the patient. Thispharmaceutical formulation with a certain concentration of theanti-IL-17 antibody must maintain physical and chemical stability of theanti-1L-17 antibody, while also avoiding viscosity that can unacceptablyincrease delivery time and force needed from a needle or auto-injectordevice.

Anti-IL-17 antibodies are disclosed in WO 07/70750 that neutralizebiological activity associated with human IL-17 (SEQ ID NO: 1). Inaddition. WO 07/70750 discloses pharmaceutical compositions ofanti-IL-17 monoclonal antibodies. With certain formulations for mAb 126,an anti-IL-17 antibody disclosed in WO 07/70750, it was discovered byApplicant as part of the present invention that three stability problemsexist at concentrations in solutions greater than or equal to 50 mg/mL:liquid-liquid phase separation; gel formation or solid phase change; andchemical instability. Thus, pharmaceutical formulations for certainconcentrations of anti-IL-17 antibodies are needed that avoid theseobserved problems. A need exists for alternative anti-IL-17 antibodypharmaceutical formulations. Further, a need exists for alternativeanti-IL-17 antibody pharmaceutical solution formulations.

Accordingly, the present invention provides a pharmaceutical formulationcomprising an anti-IL-17 antibody at a concentration in the range ofabout 80 mg/mL to about 150 mg/mL, citrate buffer at a concentration ofabout 20 mM, sodium chloride at a concentration of about 200 mM,polysorbate-80 at a concentration in the range of about 0.02% (w/v) toabout 0.03% (w/v), and a pH at about 5.7, wherein the anti-IL-17antibody comprises an antibody with a light chain (LC) and a heavy chain(HC), wherein said LC is the amino acid sequence of SEQ ID NO: 4 andsaid HC is the amino acid sequence of SEQ II) NO: 5. The presentinvention also provides a pharmaceutical formulation comprising ananti-IL-17 antibody at a concentration of about 80 mg/mL, citrate bufferat a concentration of about 20 mM, sodium chloride at a concentration ofabout 200 mM, polysorbate-80 at a concentration of about 0.03%, and pHat about 5.7, wherein the anti-IL-17 antibody comprises an antibodycomprising two light chains (LCs) and two heavy chains (HCs), whereineach LC is the amino acid sequence of SEQ ID NC): 4 and each HC is theamino acid sequence of SEQ ID NO: 5.

In addition, the present invention provides a method of treating RA, Ps,AS, PA or MM comprising administering to a patient in need thereof aneffective amount of a pharmaceutical formulation of the presentinvention. More particularly, the present invention provides a method oftreating Ps comprising administering to a patient in need thereof aneffective amount of a pharmaceutical formulation of the presentinvention. Also, the present invention provides a method of treating RAcomprising administering to a patient in need thereof an effectiveamount of a pharmaceutical formulation of the present invention. Also,the present invention provides a method of treating PA comprisingadministering to a patient in need thereof an effective amount of apharmaceutical formulation of the present invention. Also, the presentinvention provides a method of treating AS comprising administering to apatient in need thereof an effective amount of a pharmaceuticalformulation of the present invention.

In addition, the present invention provides the pharmaceuticalformulation of the present invention for use in therapy. In addition,the present invention provides the pharmaceutical formulation of thepresent invention for use in the treatment of RA, Ps, AS, PA or MM. Moreparticularly, the present invention provides the pharmaceuticalformulation of the present invention for use in the treatment of Ps.Also, the present invention provides the pharmaceutical formulation ofthe present invention for use in the treatment of RA. Also, the presentinvention provides the pharmaceutical formulation of the presentinvention for use in the treatment of PA. Also, the present inventionprovides the pharmaceutical formulation of the present invention for usein the treatment of AS.

In addition, the present invention provides the use of thepharmaceutical formulation of the present invention in the manufactureof a medicament for the treatment of RA, Ps, AS, PA or MM. Moreparticularly, the present invention provides the use of thepharmaceutical formulation of the present invention in the manufactureof a medicament for the treatment of Ps. Also, the present inventionprovides the use of the pharmaceutical formulation of the presentinvention in the manufacture of a medicament for the treatment of RA.Also, the present invention provides the use of the pharmaceuticalformulation of the present invention in the manufacture of a medicamentfor the treatment of PA. Also, the present invention provides the use ofthe pharmaceutical formulation of the present invention in themanufacture of a medicament for the treatment of AS.

Certain pharmaceutical formulations are preferred. The followingenumerated selections describe such preferred classes:

-   -   1.) the anti-IL-17 antibody comprises an antibody with an LCVR        and an HCVR, wherein said LCVR is the amino acid sequence of SEQ        ID NO: 2 and said HCVR is the amino acid sequence of SEQ ID NO:        3; comprises an antibody with a light chain (LC) and a heavy        chain (HC), wherein said LC is the amino acid sequence of SEQ ID        NO: 4 and said HC is the amino acid sequence of SEQ ID NO: 5; or        comprises an antibody comprising two light chains (LCs) and two        heavy chains (HCs), wherein each LC is the amino acid sequence        of SEQ II) NO: 4 and each HC is the amino acid sequence of SEQ        ID NO: 5;    -   2.) the anti-IL-17 antibody comprises an antibody with a light        chain (LC) and a heavy chain (HC), wherein said LC is the amino        acid sequence of SEQ ID NO: 4 and said HC is the amino acid        sequence of SEQ ID NO: 5; 3.) the anti-IL-17 antibody comprises        an antibody comprising two light chains (LCs) and two heavy        chains (HCs), wherein each LC is the amino acid sequence of SEQ        ID NO: 4 and each HC is the amino acid sequence of SEQ ID NO: 5;    -   4.) the anti-IL-17 antibody at a concentration in the range of        about 80 mg/mL to about 150 mg/mL;    -   5.) the anti-IL-17 antibody at a concentration of about 80        mg/mL;    -   6.) the polysorbate-80 at a concentration in the range of about        0.02% (w/v) to about 0.03% (w/v);    -   7.) the polysorbate-80 at a concentration of about 0.03% (w/v).

Certain pharmaceutical solution formulations are preferred. Thefollowing enumerated selections describe such preferred classes:

-   -   1.) the anti-IL-17 antibody comprises an antibody with an LCVR        and an HCVR, wherein said LCVR is the amino acid sequence of SEQ        ID NO: 2 and said HCVR is the amino acid sequence of SEQ ID NO:        3; comprises an antibody with a light chain (LC) and a heavy        chain (HC), wherein said LC is the amino acid sequence of SEQ ID        NO: 4 and said HC is the amino acid sequence of SEQ ID NO: 5; or        comprises an antibody comprising two light chains (LCs) and two        heavy chains (HCs), wherein each LC is the amino acid sequence        of SEQ ID NO: 4 and each HC is the amino acid sequence of SEQ ID        NO: 5;    -   2.) the anti-IL-17 antibody comprises an antibody with a light        chain (LC) and a heavy chain (HC), wherein said LC is the amino        acid sequence of SEQ ID NO: 4 and said HC is the amino acid        sequence of SEQ ID NO: 5;    -   3.) the anti-IL-17 antibody comprises an antibody comprising two        light chains (LCs) and two heavy chains (HCs), wherein each LC        is the amino acid sequence of SEQ ID NO: 4 and each HC is the        amino acid sequence of SEQ ID NO: 5;    -   4.) the anti-IL-17 antibody at a concentration in the range of        about 80 mg/mL to about 150 mg/mL;    -   5.) the anti-IL-17 antibody at a concentration of about 80        mg/mL;    -   6.) the polysorbate-80 at a concentration in the range of about        0.02% (w/v) to about 0.03% (w/v);    -   7.) the polysorbate-80 at a concentration of about 0.03% (w/v).

In an embodiment, the present invention also provides a pharmaceuticalformulation that comprises a concentration of anti-IL-17 antibody ofabout 80 mg/mL to about 150 mg/mL. In another embodiment, the presentinvention provides a pharmaceutical formulation that comprises aconcentration of anti-IL-17 antibody in the range of 68 mg/nit to 92mg/mL. In another embodiment, the present invention provides apharmaceutical formulation that comprises a concentration of anti-IL-17antibody of about 80 mg/mL. In another embodiment, the present inventionprovides a pharmaceutical formulation that comprises a concentration ofanti-IL-17 antibody of about 120 mg/mL. In another embodiment, thepresent invention provides a pharmaceutical formulation that comprises aconcentration of anti-IL-17 antibody of about 150 mg/mL.

In an embodiment, the present invention also provides a pharmaceuticalformulation that is buffered with citrate buffer in the range of about15 mM to about 25 mM. In another embodiment, the present inventionprovides a pharmaceutical formulation that is buffered with citratebuffer in the range of 15 mM to 25 mM. In another embodiment, thepresent invention provides a pharmaceutical formulation that is bufferedwith citrate buffer at a concentration of about 15 mM, about 20 mM,about 25 mM, or about 30 mM. In a further embodiment, the presentinvention provides a pharmaceutical formulation that is buffered withcitrate buffer at a concentration of about 20 mM.

In an embodiment, the present invention also provides a pharmaceuticalformulation that comprises a concentration of NaCl of about 200 mM toabout 300 mM. In another embodiment, the present invention provides apharmaceutical formulation that comprises a concentration of NaCl of 175mM to 225 mM. In another embodiment, the present invention provides apharmaceutical formulation that comprises a concentration of NaCl ofabout 200 mM, about 250 mM, or about 300 mM. In a further embodiment,the present invention provides a pharmaceutical formulation thatcomprises a concentration of NaCl of about 200 mM.

In an embodiment, the present invention also provides a pharmaceuticalformulation that comprises a concentration of polysorbate-80 orpolysorbate-20 of about 0.01% to about 0.04%. In another embodiment, thepresent invention provides a pharmaceutical formulation that comprises aconcentration of polysorbate-80 or polysorbate-20 of 0.02% to 0.04%. Inanother embodiment, the present invention provides a pharmaceuticalformulation that comprises a concentration of polysorbate-80 orpolysorbate-20 of about 0.01%, about 0.02%, about 0.03%, or about 0.04%.In a further embodiment, the present invention provides a pharmaceuticalformulation that comprises a concentration of polysorbate-80 orpolysorbate-20 of about 0.03%.

In an embodiment, the present invention also provides a pharmaceuticalformulation that comprises a pH range of about 5.4 to about 6.0. Inanother embodiment, the present invention provides a pharmaceuticalformulation that comprises a pH in the range of 5.4 to 6.0, in anotherembodiment, the present invention provides a pharmaceutical formulationthat comprises a pH of about 5.4, about 5.7, or about 6.0. In a furtherembodiment, the present invention provides a pharmaceutical formulationthat comprises a pH of about 5.7.

The pharmaceutical formulations of the present invention comprisecitrate buffer. Citrate buffer can be made with citric acid, trisodiumcitrate dihydrate, and citric acid monohydrate; or citric acidmonohydrate, sodium phosphate dibasic, and citric acid. Also, citratebuffer can be made comprising sodium citrate monobasic, citric acidtrisodium salt, or sodium citrate tribasic hydrate. Preferably, citratebuffer is made with sodium citrate dihydrate and citric acid.

The mAb 126 antibody is an anti-IL-17 antibody that consists of twolight chains (LCs) and two heavy chains (HCs), wherein each LC is theamino acid sequence of SEQ ID NO: 4 and each HC is the amino acidsequence of SEQ ID NO: 5, and wherein the HCs are cross-linked bydisulfide bonds.

The general structure of an “antibody” is very well-known in the art.For an antibody of the IgG type, there are four amino acid chains (two“heavy” chains and two “light” chains) that are cross-linked via intra-and inter-chain disulfide bonds. When expressed in certain biologicalsystems, antibodies having unmodified human Fc sequences areglycosylated in the Fc region. Antibodies may be glycosylated at otherpositions as well. One of skill in the art will appreciate thatantibodies for use in the formulation of the present invention maycontain such glycosylation. The subunit structures and three-dimensionalconfigurations of antibodies are well known in the art. Each heavy chainis comprised of an N-terminal heavy chain variable region (“HCVR”) and aheavy chain constant region (“HCCR”). The heavy chain constant region iscomprised of three domains (CH1, CH2, and CH3) for IgG, IgD, and IgA;and 4 domains (CH1, CH2, CH3, and CH4) for IgM and IgE. Each light chainis comprised of a light chain variable region (“LCVR”) and a light chainconstant region (“LCCR”). The variable regions of each light/heavy chainpair form the antibody binding site.

An anti-IL-17 antibody for use in the formulations of the presentinvention can be produced using techniques well known in the art, e.g.,recombinant technologies, phage display technologies, synthetictechnologies, or combinations of such technologies or other technologiesreadily known in the art. Methods for producing and purifying antibodiesand antigen-binding fragments are well known in the art and can befound, for example, in Harlow and Lane (1988) Antibodies, A LaboratoryManual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NewYork, chapters 5-8 and 15, ISBN 0-87969-314-2.

An anti-IL-17 antibody for use in the formulations of the presentinvention is an engineered antibody that has been designed to haveframeworks, hinge regions, and constant regions of human origin that areidentical with or substantially identical (substantially human) withframeworks and constant regions derived from human genomic sequences.Fully human frameworks, hinge regions, and constant regions are thosehuman germline sequences as well as sequences with naturally-occurringsomatic mutations and/or those with engineered mutations. An antibodyfor use in a formulation of the present invention may compriseframework, hinge, or constant regions derived from a fully humanframework, hinge, or constant region containing one or more amino acidsubstitutions, deletions, or additions therein. Further, an antibody foruse in a formulation of the present invention is substantiallynon-immunogenic in humans.

Stability of an antibody in solution depends on the chemical stabilityand physical stability of the antibody in the formulation in which theantibody is solubilized. Oxidation, deamidation, and hydrolysis areexamples of chemical stability issues that an antibody can have in aformulation. Aggregation and gel formation are examples of physicalstability issues that an antibody can have in a formulation. Anotherphysical stability issue with antibodies in a solution formulation maybe liquid-liquid phase separation (LLPS); LLPS in an antibody solutiontypically first appears as opalescence, followed by separation intolight and heavy phases. Cloud point is the temperature where LLPS isfirst observed for a given condition; cloud point measures thetemperature where solutions become white opaque as a result of theformation of micro-phases which ultimately resolve into the heavy andlight phases traditionally associated with phase separation.

A pharmaceutical formulation is a stable formulation wherein the degreeof degradation, modification, aggregation, loss of biological activityand the like, of proteins/antibodies therein is acceptably controlled,and does not increase unacceptably with time. Stability may be assessedby methods well-known in the art, including measurement of a sample'slight scattering, apparent attenuation of light (absorbance, or opticaldensity), size (e.g. by size exclusion chromatography (SEC)), in vitroor in vivo biological activity and/or properties measured bydifferential scanning calorimetry (DSC). Other methods for assessingstability are well known in the art and can also be used according tothe present invention. As measured by SEC, the % monomer for ananti-IL-17 pharmaceutical formulation should be greater than 90% afterstorage at 5° C. for a period of 3 months, 6 months, 9 months, 12months, 18 months, or preferably 2 years. As measured by cation exchangechromatography (CEX), the percent total acid variants should not exceed30% for an anti-Th-17 pharmaceutical formulation after storage at 5° C.for a period of 3 months, 6 months, 9 months, 12 months, 18 months, orpreferably 2 years. For measurements of purity by reduced CE-SDS, ananti-IL-17 pharmaceutical formulation should have a purity greater than85% after storage at 5° C. for a period of 3 months, 6 months, 9 months,12 months, 18 months, or preferably 2 years. Preferably, an anti-IL-17antibody pharmaceutical formulation meets one of the aforementionedstandards for stability at a temperature of 5° C. for a solution storedfor two years. More preferably, an anti-IL-17 antibody pharmaceuticalformulation meets all the aforementioned standards for stability at atemperature of 5° C. for a solution stored for two years.

The pharmaceutical formulations of the present invention can be in theliquid dosage form of a solution, emulsion, or suspension. Preferably,the pharmaceutical formulations of the present invention are in theliquid dosage form of a solution.

Administration of the pharmaceutical formulations of the presentinvention may be via parenteral administration. Parenteraladministration is commonly understood in the medical literature as theinjection of a dosage form into the body by a sterile syringe or someother mechanical device such as an infusion pump. Parenteral routes caninclude intravenous, intramuscular, subcutaneous, and intraperitonealroutes of administration. Subcutaneous administration is a preferredroute.

The pharmaceutical formulations of the present invention can be used totreat subjects with RA, Ps, AS, PA or MM. An effective amount of theanti-IL-17 antibody formulation of the present invention is the quantitywhich delivers an amount of the anti-IL-17 antibody that results in adesired therapeutic and/or prophylactic effect without causingunacceptable side-effects when administered to a subject with increasedIL-17 levels.

EXAMPLE 1 Formulating mAb 126

TABLE 1 mAb 126 Drug Product Formulation Concentration Component (mg/mL)mAb 126 80 Sodium Citrate Dihydrate 5.106 Citric Acid Anhydrous 0.507Sodium Chloride 11.69 Polysorbate 80 0.30 Water for Injection q.s. to 1mL Hydrochloric Acid pH adjustment Sodium Hydroxide pH adjustment

TABLE 2 Buffer Excipient Composition Component mg/mL Sodium CitrateDihydrate 5.106 Citric Acid Anhydrous 0.507 Sodium Chloride 11.69Polysorbate 80 0.414

The manufacturing process for the anti-IL-17 antibody pharmaceuticalsolution formulation for mAb 126 (Table 1) consists of first compoundingof the buffer excipient composition (Table 2), followed by compoundingof the final drug product formulation.

The buffer excipient composition (Table 2) is prepared, filtered, andstored for drug product formulation compounding. An appropriate quantityof water at a temperature of 20+/−5° C. is weighed into a tared emptyvessel of appropriate size. The appropriate quantity of sodium citrateis added and mixed; the appropriate quantity of citric acid is thenadded and mixed. Then, the appropriate quantity of sodium chloride isadded and mixed. Polysorbate 80 is accurately weighed out in a glasscontainer and an appropriate quantity of water at a temperature of20+/−5° C. is added into the glass container to give the concentrationin Table 2, and the solution is mixed. The entire content of thepolysorbate 80 solution is added to the other excipients. Thepolysorbate 80 solution container is rinsed with water to ensure theentire content is transferred. After addition of the polysorbate 80solution, the solution is mixed. After dissolution and mixing has beencompleted, the pH of the solution is checked to be within 5.7+/−0.2;adjustment with HCl or NaOH solution is done if necessary. The bufferexcipient composition is passed through a filter (polyvinylidenefluoride [PVDF]) for biohurden reduction.

The buffer excipient composition is supplemented with additionalpolysorbate 80 to account for the lower concentration (0.2 mg/mLpolysorbate-80) in the active pharmaceutical ingredient (API) comparedto the final drug product formulation. Since the concentration of mAb126 in the API will vary, the amount of buffer needed for dilution willalso change. This variation requires that the concentration ofpolysorbate 80 be adjusted for the buffer excipient composition recipewith each batch.

The stored mAb 126 API containers (mAb 126 is expressed in cells,purified, and concentrated; the resulting API is then frozen at about150 mg/mL to about 160 mg/mL mAb 126 in 20 mM citrate buffer, 200 mMNaCl, 0.02% polysorbate-80, at pH about 5.7) are equilibrated to atemperature of 20+/−5° C. The API solution is then mixed with anappropriate amount of the buffer excipient solution to achieve 80% ofthe theoretical batch size. The pH of the solution is checked to bewithin 5.7 +/−0.2. After pH adjustment, the solution is mixed and asample is taken for an in-process UV assay to determine the mAb 126concentration. An appropriate quantity of the buffer excipient solutionis added to reach the final target batch weight. After mixing, the pH ofthe solution is checked to be within 5.7 +/− 0.2, and the osmolality ofthe solution is checked to be within 360-480 mOsm/Kg. The mAb 126 drugproduct solution is passed through a PVDF filter for bioburden reductionand stored at 5° C.

EXAMPLE 2 Phase Separation

mAb 12.6 is found to have a propensity to phase separate while insolutions that are below 0° C. Liquid-liquid phase separation (LLPS)needs to be solved since storage of the mAb 126 drug product will be at5° C. Storage of drug product at 5° C. requires stability for periodicrefrigerator temperature excursions below 0° C. Increasing the NaClconcentration is shown to lower the temperature at which LLPS occurs inmAb 126 formulations, and increasing the citrate concentration is alsoshown to lower temperature at which LLPS occurs in mAb 126 formulations.

LLPS events are tested based on a technique developed specifically forthe temperature range (phase separation occurring between −12 and 0° C.)encountered for mAb 126. Two milliliter (mL) samples of 10 to 200 mg/mLmAb 126 were placed in a LyoStar II (FTS Systems) lyophilization unitwith the cycle shown in Table 3. Pressure was kept at atmospheric forthe experiments. Sample conditions are tested at least three times andthe standard deviation is approximately 0.5° C. for samples measured intriplicate. The samples are visually monitored during the lyophilizationcycle for signs of phase separation, including cloud point (white opaqueappearance) and the formation of a dense, protein rich layer at thebottom of the vial. Samples appear increasingly opalescent duringcooling, but this effect is easily differentiable from cloud point. Whencloud point occurs, the sample becomes an opaque white solution in lessthan a second versus a gradual increased opalescence where an objectbehind a vial is still visible through the sample.

TABLE 3 Lyophilization Cycle for Liquid-Liquid Phase Separation TestingStep 1 2 3 Cooling Rate 5 1 5 (° C./min) Target (° C.) 5 −30 20 HoldTime (min) 10 20 >20

As shown in Table 4, increasing the NaCl concentration is shown todepress the cloud point temperature. The factors which limit increasingthe NaCl concentration in the anti-IL-17 antibody pharmaceuticalformulation are hyper-tonicity and other non-phase-separation effects;200 mM NaCl avoids these problems while still depressing the cloud pointtemperature. Also, LLPS in mAb126 is shown to be bimodal at all NaClconcentrations, except 300 mM NaCl; LLPS is strongest at a concentrationof 100 mg/mL mAb 126, and the phase separation effects lessen as theconcentration moves higher or lower than 100 mg/mL.

TABLE 4 Liquid-Liquid Phase Separation: NaCl and mAb 126 ConcentrationEffects (Cloud Point ° C.) mAb 126 Conc. (mg/mL) 10 50 100 150 200  50mM NaCl −4.1 2.1 5 100 mM NaCl −1.2 2.7 1.1 150 mM NaCl −7.5 −4 −5.7−10.4 200 mM NaCl −8.5 −7 −10.2 −10.8 250 mM NaCl −6.8 −11.3 300 mM NaCl−12.5 −9.9 −12.4 −8.8

The effect of pH on cloud point of a mAb 126 solution is measured. Thecloud point was found to be minimized at two pHs, pH 4 and pH6. pH 6should be considered optimal over pH 4 when selecting a formulation formAb 126, because chemical instability of mAb 126 at pH 4 precludes usingthe low cloud point at pH 4. Given the gel formation at pH 6.3-6.4,shown in Example 3, for the mAb 126 pharmaceutical formulations, pH5.7+/−0.3 is preferred over pH 6+/−0.3 in order to allow for a pH rangethat remains stable over the shelf-life of the pharmaceuticalformulation.

Effects of various commonly used buffers on liquid-liquid phaseseparation are explored to understand what the optimal buffer systemwould be for mAb 126 (Table 5); citrate buffer is found to be mosteffective at reducing the temperature at which LLPS occurs. Thesestudies are conducted with 1.50 mM NaCl. Acetate buffer is comparable tocitrate buffer at pH 5 in terms of the effect on cloud point; however,chemical instability at pH 5 makes this pH condition less favorable.

The concentration of citrate is shown to positively affect LLPS (Table6). While citrate depresses cloud point up to 50 mM; citrate buffer isalso reported to cause more pain on injection. As such, citrateconcentrations higher than 30 mM are likely to be unacceptable from apatient compliance standpoint.

TABLE 5 Buffer Effects on Cloud Point Temperature with 150 mg/mL mAb 126Cloud Point Buffer mM pH (° C.) Acetate 10 5 −4.1 Citrate 10 5 −4.4Histidine 10 6 −2.2 Citrate 10 6 −8.1

TABLE 6 Liquid-Liquid Phase Separation: Citrate Concentration Effectswith 150 mg/mL mAb 126 Citrate conc. Cloud Point (° C.)  5 mM −6.3 10 mM−8.2 20 mM −10.2 30 mM −13.6 40 mM −14.6

EXAMPLE 3 Gel Formation

Biologic drug products are stored at 5° C.+/−3° C. to minimize chemicaland physical degradation over the shelf-life of a product. Events suchas thermodynamic solid phase change or gel formation are typically notacceptable, even if reversible, because they can negatively affectstability and hinder the required visual inspection of samples prior touse.

Solid phase change is observed in high concentration samples of mAb 126at pH conditions <pH 5 and >pH 7 at 5° C. This thermodynamic event isshown to be reversible by equilibrating the vials at room temperature.Consequently, samples are tested at various pH conditions and monitoredfor thermodynamic changes in order to find the phase boundaries moreprecisely. These tests are conducted by dialyzing samples into pH 7,citrate buffer at 5° C. to induce phase change. The solid material isthen dialyzed at the same temperature into conditions of interest toascertain if samples would reverse to the solution state. Equilibriumtesting method is preferable to long-term sample storage withintermittent inspections because, while a particular formulation may bethermodynamically unstable, the kinetics of solid phase change mayrequire months to occur in some cases.

Studies are conducted with 100 mg/mL or 150 mg/mL mAb 12.6 at 200 mMNaCl, 10 mM citrate buffer in 0.1 pH increments to find where theboundaries are for solid phase formation. The transition between phasesis found between pH 6.3 and 6.4 at 5° C. Because of the phase boundary,the target mM for a mAb 126 formulation is lowered from pH 6 to 5.7 toensure a stable storage window.

Two experiments with 80 mg/mL mAb 126 at 5° C. are carried out in 20 mMcitrate buffer, 200 mM NaCl, and 0.03% polysorbate-80; there is no gelformation at or below pH 6.1, while gel formation occurs above pH 6.1.These experiments indicate that the formulation with 80 mg/mL mAb 126has a window from pH 5.7+/−0.3 pH units wide that avoids gel formation.

EXAMPLE 4 Chemical Instability

For a pharmaceutical formulation to achieve stability, both physical andchemical sources of instability need to be addressed in the formulation.Chemical instability can result in degradation of the antibody.

In order to assess the effect of pH on chemical stability of mAb 126 at100 and 150 mg/mL samples of mAb 126 are analyzed for increases in total% acid variants by chromatography. The range of pH 4 to 7 is explored inhalf pH unit increments. The buffer for the study includes 10 mM citratebuffer, 150 mM NaCl and 0.02% polysorbate 80. Two mL solutions arestored in 3 mL glass vials with serum stoppers. Samples in these pHenvironments are stored at 5° C., 25° C., and 40° C. in order to moreaccurately model the temperature effect on the different forms ofdegradation. Samples are analyzed using Cation Exchange (CEX) HPLC usingLW detector and a Dionex ProPac WCX-10 column (4×250mm) using pH 6 10 mMBis Tris Propane (Mobile Phase A) and pH 9.6 10 mM Bis Tris Propane, 50mM NaCl (Mobile phase B).

By CEX, increases in total % acid variants (% AV) are the more reliableindicator of degradation for mAb 126. Vials of mAb 126 stored at 25° C.are shown by CEX to be most stable between pH 5-6 and least stable atmore alkaline pH conditions. When samples are stored at 40°C, pH 5 and5.5 are the most stable, with pH 6 appearing only slightly more stablethan the higher pH environments. These results show that the pH of thepharmaceutical solution formulation for mAb 126 should be between pH 5and pH 6.

EXAMPLE 5 Design of Experiment (DOE) Study

The DOE study uses a multivariate approach to examine the physical andchemical stability of mAb 126 solution formulations. mAb 126 solutionformulations are prepared according to Table 7. Each variable isexplored at five levels to measure any curvature which might exist inthe output response parameters or interactions between the inputvariables. The center point condition for the experiment is 20 mMcitrate, 200 mM NaCl, pH 5.7, 0.02% polysorbate 80. Three center pointsare dispersed throughout the design. independently preparing threecenter points provides an estimation of certainty in the analytical datawithout requiring all of the conditions to be prepared and analyzed induplicate or triplicate. Samples are stored at four temperatureconditions (5, 25, 30 and 40° C.). This range of temperatures allows forestimations of the activation energies for the results. Additionally,higher temperature storage enables earlier predictions of optimalformulation conditions.

A number of analytical techniques are selected to monitor chemical andphysical stability including size exclusion chromatography (SEC), cationexchange chromatography (CEX) HPLC, HIAC-based particle analysis,digital-image based particle analysis by micro-flow imaging (MFI,Protein Simple/Brightwell Model DPA 4200 with size range of 2-100 um),visual appearance, reduced and non-reduced Bioanalyzer Lab-on-a-Chip(LoC), pH, viscosity, and UV absorption (to measure protein content).

Using the data from all temperatures from the initial three monthperiod, activation energies (Ea) are calculated employing an Arrheniuskinetic model (zero or first Order). Energies are found using non-linearregression of all Runs. The model is then employed to extrapolate trendsout to 24 months at the relevant commercial storage temperature (5° C.).Zero order Arrhenius modeling is used for SEC (monomer, polymer Rel.Sub/Impurities), CEX (acid variants), reduced/non-reduced LoC, and UVcontent. The best fit for CEX basic variant trends is a first ordermodel.

TABLE 7 Experimental Design Run pH [PS80] [NaCl] [mAb 126] [Buffer]Buffer 1 5.7 0.02 200 120 20 Citrate 2 6 0.01 150 105 25 Citrate 3 60.03 150 105 15 Citrate 4 5.7 0 200 120 20 Citrate 5 5.4 0.03 150 135 15Citrate 6 6 0.01 250 105 15 Citrate 7 5.7 0.02 200 90 20 Citrate 8 5.70.02 100 120 20 Citrate 9 6.3 0.02 200 120 20 Citrate 10 5.1 0.02 200120 20 Citrate 11 6 0.03 250 105 25 Citrate 12 5.4 0.01 150 105 15Citrate 13 5.4 0.03 250 135 25 Citrate 14 6 0.03 150 135 25 Citrate 155.7 0.02 200 120 20 Citrate 16 5.7 0.02 200 120 10 Citrate 17 5.4 0.01250 135 15 Citrate 18 5.4 0.01 150 135 25 Citrate 19 6 0.03 250 135 15Citrate 20 5.7 0.04 200 120 20 Citrate 21 5.4 0.03 250 105 15 Citrate 226 0.01 150 135 15 Citrate 23 5.4 0.03 150 105 25 Citrate 24 5.7 0.02 300120 20 Citrate 25 5.7 0.02 200 150 20 Citrate 26 6 0.01 250 135 25Citrate 27 5.4 0.01 250 105 25 Citrate 28 5.7 0.02 200 120 30 Citrate 295.7 0.02 200 120 20 Citrate

Size Exclusion Chromatography

The two variables with the strongest effect on percent monomer based onSEC results are pH and mAb 126 concentration. The effect of mAb 126concentration is linear with increasing protein concentration resultingin decreased monomer purity.

Three input variables (pH, NaCl concentration, and buffer concentration)show curvature in their effect on percent monomer. With respect to NaCland citrate concentration, values near the center point are the moststable. Lower pH conditions are slightly more stable than the centerpoint, but other degradation pathway results make lowering the target pHless attractive.

Interactions between the effect of pH and protein concentration occurwith the effects of protein concentration being reduced at pH conditionsbelow 5.7. The two year prediction for monomer purity at the centerpoint conditions of the study is slightly below 96.7%. Polysorbate 80concentration has little effect on stability from 0.02-0.04%.

Cation Exchange Chromatography

The CEX analysis focuses on the growth of acidic species over time.Statistical modeling of CEX % AV (Ea is 23.7 kcal/g-mol) shows thatlittle chemical modification should be expected after 24 months ofstorage at 5° C. Acidic variant generation is minimized near the centerpoint for pH, but increases with greater citrate concentration. mAb 126and Polysorbate 80 concentration trends suggest that the center point isclose to the least optimal position; however, based on the magnitude ofthe y-axis scale, the difference in center point stability to otherconditions is essentially negligible.

Reduced Bioanalyzer LoC

Reduced LoC percent purity is a combination of the relative percentagesof heavy and light chains. The two input variables with the strongestinfluence on the 24-month predictions are pH and NaCl concentration.Percent purity is maximized near the center point of 200 mM NaCl. Thepercent purity increases with increasing pH. Even at the extremeformulation conditions tested in the DOE study, the molecule isstill >98% pure, indicating that the antibody is stable as measured byreduced LoC over the range of the multivariate analysis. The Ea is 21.8kcal/g-mol.

Combined Projections

All conditions in the design space studied during this experiment have 2year shelf life projections that predict <5% degradation. However, otherphysical factors preclude pH conditions above pH 6.3; therefore, thetarget conditions for the formulation should not be near this pH edge.Additionally, it is important to select a target that is in an optimalglobal maximum for the input variables explored. Based on manufacturingconsiderations, which indicate that >0.01% polysorbate 80 is needed forpumping, the polysorbate 80 target is 0.03%. Based on the results ofthis study, the optimal formulation conditions is 20 mM citrate, 200 mMNaCl. pH 5.7 with 0.03% polysorbate 80.

EXAMPLE 6

Stability at 80 mg/mL mAb 126

The stability of 80 mg/mL mAb 126 in 20 mM citrate, 200 mM NaCl, pH 5.7with 0.03% polysorbate 80 is tested out to 24 months. For storage at 5°C., stability of the anti-IL-17 antibody pharmaceutical formulation ismeasured at 0, 1, 3, 6, 9, 12, 18, and 24 months. 5° C. will be theexpected storage temperature for the anti-IL-17 antibody pharmaceuticalformulation. Accelerated stability studies at 25° C. are run for 1, 3,and 6 months.

A number of analytical techniques are selected to monitor chemical andphysical stability including size exclusion chromatography HPLC, cationexchange chromatography HPLC, visual appearance, pH, and UV absorption.CE-SDS is performed utilizing the Beckman Coulter IgGPurity/Heterogeneity kit with a Beckman Coulter ProteomeLab PA800Enhanced or Plus capillary electrophoresis (CE) instrument. For ReducedCE-SDS, samples are analyzed in a bare-fused silica capillary underdenatured, reducing conditions by molecular sieving through areplaceable gel polymer matrix after each sample injection. Fornon-reduced CE-SDS, samples are diluted to approximately 5 mg/mL inwater and subsequently diluted in sample diluent (20 mM IAM in 100 mMTris, 1% SDS, pH 9.0) to approximately 1 mg/mL. Afterwards samples areanalyzed in a bare-fused silica capillary under denatured, non-reducingconditions by molecular sieving through a replaceable gel polymer matrixat constant voltage. For both methods, UV detection is performed at 214nm. Results are shown in Table 8.

TABLE 8 Stability Data for mAb 126 at 80 mg/mL Storage Month AnalyticalProperty Condition 0 1 3 6 9 Potency (Bioassay), % 5° C. 87 — — — 108  25° C./60% RH 106   — 109 — Quantity (UV), mg/mL 5° C. 75.4 — 76.2 75.575.4 25° C./60% RH 75.3 76.0 76.5 — Monomer Purity (SEC), % 5° C. 98.3 —98.1 98.3 97.9 25° C./60% RH 98.3 97.6 97.6 — Rel Subs/Impurities: Total(SEC), % 5° C. 1.7 — 1.9 1.7  2.1 25° C./60% RH  1.7 2.4 2.4 — mAb 126Purity (CE-SDS, Reduced), % 5° C 97.8 — 97.4 97.3 97.2 25° C./60% RH97.4 96.9 96.3 — Charge Heterogeneity (CEX) [Main Peak], % 5° C. 53.9 —53.0 54.1 55.1 25° C./60% RH 56.7 59.7 60.0 — Charge Heterogeneity (CEX)[Acidic Variants], % 5° C. 13.4 — 15.4 15.3 16   25° C./60% RH 15.8 21.427.4 — Charge Heterogeneity (CEX) [Basic Variants], % 5° C. 32.8 — 31.630.7 28.9 25° C./60% RH 27.5 18.8 13.0 — pH 5° C. 5.7 — 5.7 5.7  5.7 25°C./60% RH  5.7 5.7 5.7 — Physical Appearance 5° C. NT Pass¹ — — Pass 25°C./60% RH NT — — — Particulate Matter (greater than or equal to 10 5° C.289 — 131 184 63   micrometer), particles per container 25° C./60% RH194   177 369 — Particulate Matter (greater than or equal to 25 5° C. 9— 8 31 3  micrometer), particles per container 25° C./60% RH 4  60 20 —¹Physical appearance was not performed at lot release or the 1 monthtime point. Result shown was obtained after storage at 5° C. forapproximately 2.5 months.

1-10. (canceled)
 11. A pharmaceutical formulation comprising ananti-IL-17 antibody at a concentration in the range of about 80 mg/ml toabout 150 mg/ml, citrate buffer at a concentration of about 20 mM,sodium chloride at a concentration of about 200 mM, polysorbate-80 at aconcentration in the range of about 0.02% (w/v) to about 0.03% (w/v),and pH at about 5.7, wherein the anti-IL-17 antibody comprises a lightchain variable region (LCVR) given by the amino acid sequence of SEQ IDNO:2, and a heavy chain variable region (HCVR) given by the amino acidsequence of SEQ ID NO:3, wherein LCVR comprises LCDR1, LCDR2, LCDR3, andHCVR comprises HCDR1, HCDR2, HCDR3, and wherein: (i) LCDR1 is amino acidresidues 24-39 of SEQ ID NO: 2, and the amino acid residue at position 4is Gln, Ala, Lys, or Arg, the amino acid residue at position 10 is anArg, His, Asn, Tyr, Ala, or Leu, and the amino acid residue at position14 is Tyr, Phe, or Trp; (ii) LCDR2 is amino acid residues 55-61 of SEQID NO: 2, and the amino acid residue at position 3 is Ser, His, Pro,Arg, Asp, or Ala, the amino acid residue at position 7 is Ser, Ile, His,Val, or Thr; (iii) LCDR3 is amino acid residues 94-102 of SEQ ID NO: 2,and the amino acid residue at position 3 is Ser or Thr, the amino acidresidue at position 4 is Thr or Met, the amino acid residue at position6 is Leu, Tyr, Ile, Val, or Glu; (iv) HCDR1 is amino acid residues 26-35of SEQ ID NO: 3, and the amino acid residue at position 8 is Asn, His,the amino acid residue at position 9 is Met, Ile, Leu, Thr, the aminoacid residue at position 10 is Asn, Ser, Gly, or His; (v) HCDR2 is aminoacid residues 50-66 of SEQ ID NO: 3, and the amino acid residue atposition 5 is Asn, Met, Ala, or Glu; and (vi) HCDR3 is amino acidresidues 99-108 of SEQ ID NO: 3, and the amino acid residue at position4 is Ala, Trp, His, Phe, Tyr, Ser, Leu, the amino acid residue atposition 9 is Ala, Gly, Leu, Val, Thr, or Pro.
 12. The formulation ofclaim 11, wherein Arg is in position 4, Arg is in position 10, and Tyris in position 14 of LCDR1.
 13. The formulation of claim 11, wherein Seris in position 3 and Ile is in position 7 of LCDR2.
 14. The formulationof claim 11, wherein Ser is in position 3, Thr is in position 4, and Leuis in position 6 of LCDR3.
 15. The formulation of claim 11, wherein Hisis in position 8, Ile is in position 9, and His is in position 10 ofHCDR1.
 16. The formulation of claim 11, wherein Met is in position 5 ofHCDR2.
 17. The formulation of claim 11, wherein Phe is in position 4 andVal is in position 9 of HCDR3.
 18. The formulation of claim 11, whereinArg is in position 4, Arg is in position 10, and Tyr is in position 14of LCDR1, Ser is in position 3 and Ile is in position 7 of LCDR2, Ser isin position 3, Thr is in position 4, and Leu is in position 6 of LCDR3,His is in position 8, Ile is in position 9, and His is in position 10 ofHCDR1, Met is in position 5 of HCDR2, Phe is in position 4 and Val is inposition 9 of HCDR3.
 19. The formulation of claim 11, wherein theconcentration of the anti-IL-17 antibody is about 80 mg/ml.
 20. Theformulation of claim 18, wherein the concentration of the anti-IL-17antibody is about 80 mg/ml.
 21. The formulation of claim 19, wherein theformulation is an anti-IL-17 antibody pharmaceutical solutionformulation.
 22. The formulation of claim 20, wherein the formulation isan anti-IL-17 antibody pharmaceutical solution formulation.
 23. A methodof treating rheumatoid arthritis, psoriasis, ankylosing spondilitis,psoriatic arthritis or multiple myeloma comprising administering to apatient in need thereof an effective amount of the pharmaceuticalformulation of claim
 11. 24. A method of treating rheumatoid arthritis,psoriasis, ankylosing spondilitis, psoriatic arthritis or multiplemyeloma comprising administering to a patient in need thereof aneffective amount of the pharmaceutical formulation of claim
 18. 25. Amethod of treating rheumatoid arthritis, psoriasis, ankylosingspondilitis, psoriatic arthritis or multiple myeloma comprisingadministering to a patient in need thereof an effective amount of thepharmaceutical solution formulation of claim
 19. 26. A method oftreating rheumatoid arthritis, psoriasis, ankylosing spondilitis,psoriatic arthritis or multiple myeloma comprising administering to apatient in need thereof an effective amount of the pharmaceuticalsolution formulation of claim
 20. 27. A method of treating rheumatoidarthritis, psoriasis, ankylosing spondilitis, psoriatic arthritis ormultiple myeloma comprising administering to a patient in need thereofan effective amount of the pharmaceutical solution formulation of claim21.
 28. A method of treating rheumatoid arthritis, psoriasis, ankylosingspondilitis, psoriatic arthritis or multiple myeloma comprisingadministering to a patient in need thereof an effective amount of thepharmaceutical solution formulation of claim 22.